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Studies on North-West Himalayan Russulaceae

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Studies on North-West Himalayan Russulaceae Biological Forum – An International Journal 13(2): 138-164(2021) ISSN No. (Print): 0975-1130 ISSN No. (Online): 2249-3239 Studies on North-West Himalayan Russulaceae Shilpa Sood1*, Reeti Singh2 and Ramesh Chandra Upadhyay3 1Department of Botany, MCM DAV College Kangra, (Himachal Pradesh)-176001, India. 2Department of Plant Pathology, College of Agriculture, RVSK Vishwa Vidyalaya, Gwalior, (Madhya Pradesh)-474002, India. 3ICAR-Directorate of Mushroom Research, Solan, (Himachal Pradesh)-173213, India. (Corresponding author: Shilpa Sood*) (Received 22 March 2021, Accepted 05 June, 2021) (Published by Research Trend, Website: www.researchtrend.net) ABSTRACT: North Western Himalayan forests are rich in macrofungal diversity, especially in ectomycorrhizal (ECM) fungi. In 2014 and 2015, a number of explorations were undertaken during rainy season to explore the ECM diversity. The morpho-taxonomy of thirteen samples of Russulaceae were briefly discussed. Four samples of Lactarius and nine samples of Russula were described morphologically and illustrated taxonomically. Detailed study on the spore morphology of the specimens was carried out using staining techniques and Scanning Electron Microscopy (SEM). Out of these Lactarius paradoxus, L. subpurpureus, Russula fellea, R. flavida and R. subfoetens have been reported for the first time from Himachal Pradesh. Keywords: Basidiomycetes, macrofungi, Russula, Lactarius, taxonomy. INTRODUCTION and injury, latex colour and latex colour change were described in the field from the fresh specimen. The members of family Russulaceae belongs to All colour notations were according to Kornerup and cosmopolitan group of ectomycorrhizal mushroom that Wanscher, (1978). After recording all the forms a relationship with trees. It comprises around morphological characters, specimens were dried in hot 1900 accepted species (Lebel et al., 2013). Major air oven and preserved in polypropylene bags with genera of this group are Russula, Multifurca, Lactarius naphthalene ball and some crystals of 2,4- and Lactifluus are easily distinguished from other gilled dichlorobenzene. Microscopic studies were done from mushrooms by the consistency of their flesh, which is dried sample revived in 3% KOH. Hymenium, basidia, granular, brittle, breaks easily and absence of volva. cystidia, pileipellis and stipitipellis were observed in Lactarius consists of 450 spp. worldwide (Kirk et al., 1% congo red and 1% cotton blue under oil immersion 2008) and 83 from India (Upadhyay et al., 2017). (Motic BA 310). Spores were studied from the spore Russula consists of 750 spp. worldwide (Kirk et al., deposits in Melzer’s reagent. Camera lucida drawings 2008) and 128 spp. from India (Upadhyay et al., 2017). were made with the aid of a drawing tube. Spore In the present study four species of Lactarius: L. ornamentations were observed by SEM using camphorates, L. paradoxus, L. salmoneus, L. EMCRAFT microscope. The examined collections subperpureus and nine species of Russula: R. were deposited in the Herbarium of ICAR, Directorate atropurpurea, R. brevipes var. acrior, R. delica, R. of Mushroom Research, Chambaghat, Solan, (H.P.). flavida, R. fellea, R. foetens, R. fragilis, R. grisea, R. subfoetens were reported from forests of Himachal RESULTS Pradesh. Two Lactarius (L. paradoxus and L. Lactarius camphoratus (Bull.) Fr., Epicrisis Systematis subpurpureus) and three Russula (R. fellea, R. flavida Mycologici: 346, 1838 [Figs. 1 (A H) and 2 (A F)] and R. subfoetens) are first time documented from – – Himachal Pradesh. Pileus diameter up to 4.0 cm wide, umbonate, greyish orange (6B-5) to (6B-4), surface non hygrophanous, MATERIALS AND METHODS margin irregular, striate, inflexed, dry, smooth, scales Specimens were collected during rainy season. absent, cuticle not peeling, pileus consistency fleshy, Specimens were photographed and all the context colour cream, up to 0.1cm thick, no colour morphological traits such as colour, shape, size and change on touch or handling and confluent pileus. change in colour on bruising of carpophores on touch Lamellae shortly decurrent, close, separable, flesh coloured (6B-3), fleshy, gill size 2.0 × 0.2 cm unequal Sood et al., Biological Forum – An International Journal 13(2): 138-164(2021) 138 present in 5 sets of lamellulae, secrete milky latex when (11.3) × (3.5)4.9–(9.7) µm wide, septate, branched, injured. Stipe central, light brown (6D-5) to brown (6E- septa without clamps. 5), cylindric 8.5 × 0.3 cm, terete, dry, glabrous, blunt at Collection examined: India- Himachal Pradesh- the base, hollow, fleshy, stipe trama colour cream, ring, Shimla- Kufri: alt. 2290 m a.s.l.; GPS 31°6'0" veil and volva absent. N77°15'0"E, mixed forest, under Pine tree. Shilpa Basidiospores (7.2)8.0–(8.8) × (5.8)6.5–(7.6) µm Sood, DMR Acc. No. 23/15, July 26, 2015. (Q=1.1−1.3), hyaline, ornamented, subglobose to Discussion. L. camphoratus has been misidentified as eliptical, apiculate (0.9–1.5 µm), germ pore absent, oil L. rubidus due to red brown pileus, close to crowded globule present, cyanophilic and amyloid. Basidia pale pinkish cinnamon lamellae. However, L. rubidus is (26.4)30.8–(35.6) × (7.4)9.4–(12.9) µm (Q=3.3), a European and Eastern North American species clavate, 2, 4-spored, basidia mostly tetra-sterigmatic, featuring watery latex and absence of pleurocystidia. L. long curbed sterigmata up to 5.5 × 1.7 µm, thin walled, camphoratus can be identified due to variable cystidia oil globule present, basal septa without clamps. from subclavate, subcylindric to fusiform, irregular and Pleurocystidia present, (33.0)38.5–(49.8) × (7.5)10.0– nearly lobed. It is also confused with L. fragilis which (12.0) µm, cheilocystidia (24.3)28.8–(35.0) × (5.4)6.8– has watery latex yellowish gills, and (more definitively) (9.0) µm. Pileipellis (10.2)14.8–(22.7) × (8.8)12.1– spores that are reticulate. This genus is already reported (17.2) µm thick, hyphae irregularly arranged, thin form Himachal Pradesh: Bhatt and Lakhanpal, (1990); walled, with some cylindrical hyphae. Hymenophoral Uttarakhand: Bhatt et al., (2000), Das and Sharma, trama (3.9)5.3–(8.8) µm thick, regular. Subhymenium (2002). 6–13 µm. Hymenium 17–23 µm. Stipe cuticle (6.4)7.9– Fig. 1 (A–H) Lactarius camphoratus: A) Basidiocarps in natural habitat; B) Basidiocarp with slight umbo; C) Gills with milky latex; D) Lobed cystidia; E) Pileipellis; F) Basidiospores in light microscope; G and H) SEM of basidiospores. Scale bars: D–F=10 µm; G=1 µm; H=2 µm. Sood et al., Biological Forum – An International Journal 13(2): 138-164(2021) 139 Fig. 2 (A–F) Lactarius camphoratus: A) Basidiocarps; B) Basidiospores; C) Basidia; D) Pleurocystidia; E) Cheilocystidia; F) Pileipellis. Scale bars: B–F=10 µm. Lactarius paradoxus Beardslee & Burl., Mycologia 32: absent, oil globule present (1–2 in no.), cyanophilic and 586, 1940 [Figs. 3 (A–F) and 4 (A–H)] amyloid. Basidia (38.2)43.4–(50.5) × (6.7)8.8–(10.7) Pileus up to 8.7 cm diam. depressed, purplish pink µm, (Q=4.9), clavate, 4-spored, sterigmata up to 6.0 × (14A-3) to reddish lilac (14B-3), regular, wavy, non- 1.9 µm, thin walled, oil globule present, basal septa striate, with concentric zones, inflexed, moist, non without clamps. Pleurocystidia present, fusiform to hygrophanous, smooth, scales absent, fleshy, colour moniliform (37.6).59.2–(70.6) × (4.6)7.8–(8.6) µm, change on touch pale blue (21A-3). Lamellae purplish cheilocystidia present (19.6)24.7–(37.6) × (3.6)5.9– white (14A-2) to purplish pink (14A-3), adnate, (8.2) µm, same as like pleurocystidia, oil globule unequal, close, separable, 7 sets of lamellulae, fleshy, present, clamp connections absent. Pileipellis ixocutis. interveined, separable, latex blood red in colour. Stipe Hymenophoral trama (3.3)8.8–(14.4) µm thick with 2.9 × 1.2 cm, central, purplish white (14A-2), cylindric, laticiferous hyphae. Subhymenium 15–22 µm. blunt, hollow, terete, fleshy, ring, veil and volva absent. Hymenium 35–45 µm. Stipe cuticle regular strongly Basidiospores (8.8)9.1–(9.3) × (5.1)6.0–(7.2) µm interwoven hyphae (1.8)2.9–(4.2) µm. (Q=1.5), hyaline, elliptical, reticulate, SEM studies Collection examined: India- Himachal Pradesh- showed ornamentation up to 0.5 μm high, of rather Shimla- Narkanda- Oddi: alt. 2621 m a.s.l; GPS thick ridges, forming an almost complete reticulum, 32°9'52"N 76°16'39"E, coniferous, gregarious, on soil isolated warts scarce, plage distinct less isolated warts among grasses. Shilpa Sood, DMR Acc. No. 53/14, and slightly amyloid, apiculate (0.8–1.4 µm), germ pore September 21, 2014. Sood et al., Biological Forum – An International Journal 13(2): 138-164(2021) 140 Discussion. L. paradoxus is a medium sized species, with differently ornamented spores. From India, Atri characterized by the bluish and often greenish tinges in and Saini, (1986) first reported this species form the pileus and the vinaceous red to brown latex. Uttarakhand. In the present investigation the specimen Remarkable is the short root-like projection of the stipe. was found scatter to gregarious in coniferous forest and Hesler and Smith, (1979) reported several collections reported first time from Himachal Pradesh. Fig. 3 (A–F) Lactarius paradoxus: A and B) Basidiocarps in natural habitat; B) Basidiospores showing zonation in pileus suface and gill attachment; C) Basidiospores in light microscope; D) Basidiospores in SEM; E) Pileipellis; F) Section showing abundant laticifer hyphae in gill trama. Scale bars: C, E and F=10 µm; D=1 µm. Sood et al., Biological Forum – An International Journal 13(2): 138-164(2021) 141 Fig. 4 (A–H) Lactarius paradoxus: A) Basidiocarps; B) Basidiospores; C) Basidia; D) Pleurocystidia; E) Cheilocystidia; F) Pseudocystidia; G) Pileipellis; H) Stipitopellis. Scale bars: B–H=10 µm.
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